Decellularization protocol-dependent damage-associated molecular patterns in rat uterus scaffolds differentially affect the immune response after transplantation

Arvind Manikantan Padma; Ahmed Baker Alshaikh; Min Jong Song; Randa Akouri; Mihai Oltean; Mats Brännström; Mats Hellström

doi:10.1002/term.3217

Decellularization protocol-dependent damage-associated molecular patterns in rat uterus scaffolds differentially affect the immune response after transplantation

Arvind Manikantan Padma, Ahmed Baker Alshaikh, Min Jong Song, Randa Akouri, Mihai Oltean, Mats Brännström, Mats Hellström

Research output: Contribution to journal › Article › peer-review

22 Scopus citations

Abstract

Scaffolds derived from decellularized tissue possess many advantages for bioengineering applications, including for novel infertility treatments. However, the decellularization process results in allogenic-independent damage-associated molecular patterns (DAMPs). This field is poorly studied, in particular for uterus bioengineering applications. An increased knowledge concerning the immune system activation after transplantation of decellularized tissue will enable safer construct development and thereby accelerate translation from research to clinic. We therefore transplanted rat uterus scaffolds produced by three different decellularization protocols based on Triton X-100 (P1 and P2) or sodium deoxycholate (P3) in a syngeneic animal model and assessed the immune response towards DAMPs exposed by the decellularization process. Biopsies were retrieved on day 5, 15, and 30 post transplantation and immunohistochemistry-stained CD45⁺ (leucocytes), CD4⁺ (T-cells), CD8a⁺ (cytotoxic T-cells), CD22⁺ (B-cells), NCR1⁺ (NK-cells), CD68⁺ (pan-macrophages), and CD163⁺ (M2 macrophages) cells within the grafts were quantified. The gene expression for interferon γ, interleukin (IL)-1β, IL-2, IL-6, and tumor necrosis factor (TNF) eotaxin-2, RANTES, MCP-1, MIP-1α, MIP-3α, IL-8 were also measured. Scaffolds from P1 induced a rapid cell infiltration after transplantation, presumably induced by DNA-based DAMPs. However, this response was only transient. Protocol 3 derived scaffolds induced an early pro-inflammatory cytokine response at the transcript level which remained high throughout the study. This response may be caused by the stronger decellularization detergent that could expose more extracellular matrix-related DAMPs. However, earlier proteomics analysis also identified significantly more abundant heat shook proteins-related DAMPs in this scaffold type. Protocol 2 caused the least immunogenic scaffolds and should thus be the future focus for in vivo uterus bioengineering applications.

Original language	English
Pages (from-to)	674-685
Number of pages	12
Journal	Journal of Tissue Engineering and Regenerative Medicine
Volume	15
Issue number	7
DOIs	https://doi.org/10.1002/term.3217
State	Published - Jul 2021

Bibliographical note

Funding Information:
We thank Prof. Levent Akürek and Prof. Suchitra Sumitran for valuable discussions. The study was financed by the Knut and Alice Wallenberg foundation, the Swedish research council (VR: 116008), the ALF-agreement (between the Swedish government and the county council), the Adlerbertska, Hjalmar Svensson, and Wilhelm & Martina Lundgrens research foundations, the Swedish-Korean International Cooperation in Research and Higher Education (STINT) and the International Research and Development Program of the NRF of Korea, funded by the Ministry of Science and ICT of Korea (NRF-2018K2A9A2A12000216 and NRF-2018R1D1A1B07049059).

Funding Information:
We thank Prof. Levent Akürek and Prof. Suchitra Sumitran for valuable discussions. The study was financed by the Knut and Alice Wallenberg foundation, the Swedish research council (VR: 116008), the ALF‐agreement (between the Swedish government and the county council), the Adlerbertska, Hjalmar Svensson, and Wilhelm & Martina Lundgrens research foundations, the Swedish‐Korean International Cooperation in Research and Higher Education (STINT) and the International Research and Development Program of the NRF of Korea, funded by the Ministry of Science and ICT of Korea (NRF‐2018K2A9A2A12000216).

Publisher Copyright:
© 2021 The Authors. Journal of Tissue Engineering and Regenerative Medicine published by John Wiley & Sons Ltd.

Keywords

bioengineering
DAMPs
decellularization
immune response
transplantation
uterus

Access to Document

10.1002/term.3217

Cite this

Padma, A. M., Alshaikh, A. B., Song, M. J., Akouri, R., Oltean, M., Brännström, M., & Hellström, M. (2021). Decellularization protocol-dependent damage-associated molecular patterns in rat uterus scaffolds differentially affect the immune response after transplantation. Journal of Tissue Engineering and Regenerative Medicine, 15(7), 674-685. https://doi.org/10.1002/term.3217

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title = "Decellularization protocol-dependent damage-associated molecular patterns in rat uterus scaffolds differentially affect the immune response after transplantation",

abstract = "Scaffolds derived from decellularized tissue possess many advantages for bioengineering applications, including for novel infertility treatments. However, the decellularization process results in allogenic-independent damage-associated molecular patterns (DAMPs). This field is poorly studied, in particular for uterus bioengineering applications. An increased knowledge concerning the immune system activation after transplantation of decellularized tissue will enable safer construct development and thereby accelerate translation from research to clinic. We therefore transplanted rat uterus scaffolds produced by three different decellularization protocols based on Triton X-100 (P1 and P2) or sodium deoxycholate (P3) in a syngeneic animal model and assessed the immune response towards DAMPs exposed by the decellularization process. Biopsies were retrieved on day 5, 15, and 30 post transplantation and immunohistochemistry-stained CD45+ (leucocytes), CD4+ (T-cells), CD8a+ (cytotoxic T-cells), CD22+ (B-cells), NCR1+ (NK-cells), CD68+ (pan-macrophages), and CD163+ (M2 macrophages) cells within the grafts were quantified. The gene expression for interferon γ, interleukin (IL)-1β, IL-2, IL-6, and tumor necrosis factor (TNF) eotaxin-2, RANTES, MCP-1, MIP-1α, MIP-3α, IL-8 were also measured. Scaffolds from P1 induced a rapid cell infiltration after transplantation, presumably induced by DNA-based DAMPs. However, this response was only transient. Protocol 3 derived scaffolds induced an early pro-inflammatory cytokine response at the transcript level which remained high throughout the study. This response may be caused by the stronger decellularization detergent that could expose more extracellular matrix-related DAMPs. However, earlier proteomics analysis also identified significantly more abundant heat shook proteins-related DAMPs in this scaffold type. Protocol 2 caused the least immunogenic scaffolds and should thus be the future focus for in vivo uterus bioengineering applications.",

keywords = "bioengineering, DAMPs, decellularization, immune response, transplantation, uterus",

author = "Padma, {Arvind Manikantan} and Alshaikh, {Ahmed Baker} and Song, {Min Jong} and Randa Akouri and Mihai Oltean and Mats Br{\"a}nnstr{\"o}m and Mats Hellstr{\"o}m",

note = "Funding Information: We thank Prof. Levent Ak{\"u}rek and Prof. Suchitra Sumitran for valuable discussions. The study was financed by the Knut and Alice Wallenberg foundation, the Swedish research council (VR: 116008), the ALF-agreement (between the Swedish government and the county council), the Adlerbertska, Hjalmar Svensson, and Wilhelm & Martina Lundgrens research foundations, the Swedish-Korean International Cooperation in Research and Higher Education (STINT) and the International Research and Development Program of the NRF of Korea, funded by the Ministry of Science and ICT of Korea (NRF-2018K2A9A2A12000216 and NRF-2018R1D1A1B07049059). Funding Information: We thank Prof. Levent Ak{\"u}rek and Prof. Suchitra Sumitran for valuable discussions. The study was financed by the Knut and Alice Wallenberg foundation, the Swedish research council (VR: 116008), the ALF‐agreement (between the Swedish government and the county council), the Adlerbertska, Hjalmar Svensson, and Wilhelm & Martina Lundgrens research foundations, the Swedish‐Korean International Cooperation in Research and Higher Education (STINT) and the International Research and Development Program of the NRF of Korea, funded by the Ministry of Science and ICT of Korea (NRF‐2018K2A9A2A12000216). Publisher Copyright: {\textcopyright} 2021 The Authors. Journal of Tissue Engineering and Regenerative Medicine published by John Wiley & Sons Ltd.",

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doi = "10.1002/term.3217",

language = "English",

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Padma, AM, Alshaikh, AB, Song, MJ, Akouri, R, Oltean, M, Brännström, M & Hellström, M 2021, 'Decellularization protocol-dependent damage-associated molecular patterns in rat uterus scaffolds differentially affect the immune response after transplantation', Journal of Tissue Engineering and Regenerative Medicine, vol. 15, no. 7, pp. 674-685. https://doi.org/10.1002/term.3217

Decellularization protocol-dependent damage-associated molecular patterns in rat uterus scaffolds differentially affect the immune response after transplantation. / Padma, Arvind Manikantan; Alshaikh, Ahmed Baker; Song, Min Jong et al.
In: Journal of Tissue Engineering and Regenerative Medicine, Vol. 15, No. 7, 07.2021, p. 674-685.

Research output: Contribution to journal › Article › peer-review

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AU - Padma, Arvind Manikantan

AU - Alshaikh, Ahmed Baker

AU - Song, Min Jong

AU - Akouri, Randa

AU - Oltean, Mihai

AU - Brännström, Mats

AU - Hellström, Mats

N1 - Funding Information: We thank Prof. Levent Akürek and Prof. Suchitra Sumitran for valuable discussions. The study was financed by the Knut and Alice Wallenberg foundation, the Swedish research council (VR: 116008), the ALF-agreement (between the Swedish government and the county council), the Adlerbertska, Hjalmar Svensson, and Wilhelm & Martina Lundgrens research foundations, the Swedish-Korean International Cooperation in Research and Higher Education (STINT) and the International Research and Development Program of the NRF of Korea, funded by the Ministry of Science and ICT of Korea (NRF-2018K2A9A2A12000216 and NRF-2018R1D1A1B07049059). Funding Information: We thank Prof. Levent Akürek and Prof. Suchitra Sumitran for valuable discussions. The study was financed by the Knut and Alice Wallenberg foundation, the Swedish research council (VR: 116008), the ALF‐agreement (between the Swedish government and the county council), the Adlerbertska, Hjalmar Svensson, and Wilhelm & Martina Lundgrens research foundations, the Swedish‐Korean International Cooperation in Research and Higher Education (STINT) and the International Research and Development Program of the NRF of Korea, funded by the Ministry of Science and ICT of Korea (NRF‐2018K2A9A2A12000216). Publisher Copyright: © 2021 The Authors. Journal of Tissue Engineering and Regenerative Medicine published by John Wiley & Sons Ltd.

PY - 2021/7

Y1 - 2021/7

N2 - Scaffolds derived from decellularized tissue possess many advantages for bioengineering applications, including for novel infertility treatments. However, the decellularization process results in allogenic-independent damage-associated molecular patterns (DAMPs). This field is poorly studied, in particular for uterus bioengineering applications. An increased knowledge concerning the immune system activation after transplantation of decellularized tissue will enable safer construct development and thereby accelerate translation from research to clinic. We therefore transplanted rat uterus scaffolds produced by three different decellularization protocols based on Triton X-100 (P1 and P2) or sodium deoxycholate (P3) in a syngeneic animal model and assessed the immune response towards DAMPs exposed by the decellularization process. Biopsies were retrieved on day 5, 15, and 30 post transplantation and immunohistochemistry-stained CD45+ (leucocytes), CD4+ (T-cells), CD8a+ (cytotoxic T-cells), CD22+ (B-cells), NCR1+ (NK-cells), CD68+ (pan-macrophages), and CD163+ (M2 macrophages) cells within the grafts were quantified. The gene expression for interferon γ, interleukin (IL)-1β, IL-2, IL-6, and tumor necrosis factor (TNF) eotaxin-2, RANTES, MCP-1, MIP-1α, MIP-3α, IL-8 were also measured. Scaffolds from P1 induced a rapid cell infiltration after transplantation, presumably induced by DNA-based DAMPs. However, this response was only transient. Protocol 3 derived scaffolds induced an early pro-inflammatory cytokine response at the transcript level which remained high throughout the study. This response may be caused by the stronger decellularization detergent that could expose more extracellular matrix-related DAMPs. However, earlier proteomics analysis also identified significantly more abundant heat shook proteins-related DAMPs in this scaffold type. Protocol 2 caused the least immunogenic scaffolds and should thus be the future focus for in vivo uterus bioengineering applications.

AB - Scaffolds derived from decellularized tissue possess many advantages for bioengineering applications, including for novel infertility treatments. However, the decellularization process results in allogenic-independent damage-associated molecular patterns (DAMPs). This field is poorly studied, in particular for uterus bioengineering applications. An increased knowledge concerning the immune system activation after transplantation of decellularized tissue will enable safer construct development and thereby accelerate translation from research to clinic. We therefore transplanted rat uterus scaffolds produced by three different decellularization protocols based on Triton X-100 (P1 and P2) or sodium deoxycholate (P3) in a syngeneic animal model and assessed the immune response towards DAMPs exposed by the decellularization process. Biopsies were retrieved on day 5, 15, and 30 post transplantation and immunohistochemistry-stained CD45+ (leucocytes), CD4+ (T-cells), CD8a+ (cytotoxic T-cells), CD22+ (B-cells), NCR1+ (NK-cells), CD68+ (pan-macrophages), and CD163+ (M2 macrophages) cells within the grafts were quantified. The gene expression for interferon γ, interleukin (IL)-1β, IL-2, IL-6, and tumor necrosis factor (TNF) eotaxin-2, RANTES, MCP-1, MIP-1α, MIP-3α, IL-8 were also measured. Scaffolds from P1 induced a rapid cell infiltration after transplantation, presumably induced by DNA-based DAMPs. However, this response was only transient. Protocol 3 derived scaffolds induced an early pro-inflammatory cytokine response at the transcript level which remained high throughout the study. This response may be caused by the stronger decellularization detergent that could expose more extracellular matrix-related DAMPs. However, earlier proteomics analysis also identified significantly more abundant heat shook proteins-related DAMPs in this scaffold type. Protocol 2 caused the least immunogenic scaffolds and should thus be the future focus for in vivo uterus bioengineering applications.

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Decellularization protocol-dependent damage-associated molecular patterns in rat uterus scaffolds differentially affect the immune response after transplantation

Abstract

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Keywords

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